Nanosphere lithography fabricated nanostructures have highly tunable localized surface plasmons, which have been used for important sensing and spectroscopy applications. In this work, the authors focus on biological applications and technologies that utilize two types of related plasmonic phenomena: localized surface plasmon resonance (LSPR) spectroscopy and surface-enhanced Raman spectroscopy (SERS). Two applications of these plasmonic materials are presented: (i) the development of an ultrasensitive nanoscale optical biosensor based on LSPR wavelength-shift spectroscopy and (ii) the SERS detection of an anthrax biomarker.